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Selective LXXLL peptides antagonize transcriptional activation by the retinoid-related orphan receptor RORgamma

Shogo Kurebayashi1, Takeshi Nakajima, Seong-Chul Kim

  • 1Cell Biology Section, Division of Intramural Research, National Institute of Environmental Sciences, National Institutes of Health, Research Triangle Park, NC 27709, USA.

Biochemical and Biophysical Research Communications
|February 27, 2004
PubMed

Insights

Retinoid-related orphan receptor gamma (RORgamma) interaction with co-activators was studied using LXXLL peptides. These peptides antagonized RORgamma activity and revealed distinct binding site conformations, aiding RORgamma signaling research.

Area of Science:

  • Molecular biology
  • Endocrinology
  • Structural biology

Background:

  • Retinoid-related orphan receptor gamma (RORgamma) is a nuclear receptor that regulates transcription.
  • Nuclear receptor transcriptional activation depends on co-activator recruitment via LXXLL motifs.
  • Understanding RORgamma's co-activator interaction is crucial for deciphering its regulatory mechanisms.

Purpose of the Study:

  • To investigate the interaction between RORgamma and the co-activator SRC1.
  • To analyze the RORgamma ligand-binding domain (LBD) co-activator interaction surface using LXXLL peptides.
  • To identify specific amino acid residues critical for LXXLL peptide binding and RORgamma transcriptional activity.

Main Methods:

  • Utilized a series of LXXLL-containing peptides to probe RORgamma LBD conformation.
  • Employed site-directed mutagenesis to alter key residues in the RORgamma LBD.
  • Performed homology modeling to predict the structure of the RORgamma LBD.

Main Results:

  • RORgamma's H3-4/H12 surface exhibits unique selectivity for LXXLL peptides compared to other nuclear receptors.
  • LXXLL peptides that bind RORgamma effectively antagonize its transcriptional activity.
  • Specific mutations (E502Q, Y500F, K334A, Q347A, I348D) significantly impair LXXLL peptide recruitment.

Conclusions:

  • The identified LXXLL peptide antagonists are valuable tools for studying RORgamma LBD conformation.
  • Mutational analysis supports homology modeling predictions of the RORgamma LBD structure.
  • This research provides insights into RORgamma receptor signaling and potential therapeutic targeting.

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